Protection-Based Assays to Measure Aminoacyl-tRNA Binding to Translation Initiation Factors

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

To decipher the mechanisms of translation initiation, the stability of the complexes between tRNA and initiation factors has to be evaluated in a routine manner. A convenient method to measure the parameters of binding of an aminoacyl-tRNA to an initiation factor results from the property that, when specifically complexed to a protein, the aminoacyl-tRNA often resists spontaneous deacylation. This chapter describes the preparation of suitable aminoacyl-tRNA ligands and their use in evaluating the stability of their complexes with various initiation factors, such as e/aIF2 and e/aIF5B. The advantages and the limitations of the method are discussed.

Original languageEnglish
Title of host publicationTranslation Initiation
Subtitle of host publicationReconstituted Systems and Biophysical Methods
PublisherAcademic Press Inc.
Pages265-281
Number of pages17
ISBN (Print)9780123739698
DOIs
Publication statusPublished - 1 Jan 2007

Publication series

NameMethods in Enzymology
Volume430
ISSN (Print)0076-6879

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